CN116249699A

CN116249699A - SGLT-2 inhibitor-sarcosine eutectic, preparation method and application thereof

Info

Publication number: CN116249699A
Application number: CN202080104942.8A
Authority: CN
Inventors: 贾慧娟; 侯伟; 王艳鑫; 李衍; 任晓慧; 贾天格
Original assignee: Beijing Creatron Institute Of Pharmaceutical Research Co ltd
Current assignee: Beijing Creatron Institute Of Pharmaceutical Research Co ltd
Priority date: 2020-09-30
Filing date: 2020-09-30
Publication date: 2023-06-09
Also published as: US20230372375A1; EP4206212A1; EP4206212A4; WO2022067724A1

Abstract

Provides an SGLT-2 inhibitor sarcosine eutectic, a preparation method and application thereof. The sarcosine is used as a ligand, so that the method has higher safety and lower cost; the pharmaceutical co-crystal has higher stability, and the crystal form is not easy to change in the production process or the storage process of the preparation composition; the mixture is not melted when being heated, and is not sticky, clustered or static, so that the mixture has better mixing uniformity; the medicine composition is uniformly distributed in the prepared medicine composition, so that the medicine composition has better in vivo release, absorption and medicine effect and small batch-to-batch difference; the product has high stability, and is more beneficial to storage and transportation; the preparation process is simple, the repeatability is high, the crystallization time is short, the process condition requirement is low, and the economic benefit is higher; unsafe solvents are not used in the preparation process; crude or intermediate SGLT-2 inhibitors may be purified simultaneously.

Description

SGLT-2 inhibitor-sarcosine eutectic, preparation method and application thereof

Technical Field

The invention relates to the technical field of chemical pharmacy, in particular to an SGLT-2 inhibitor and sarcosine eutectic, and a preparation method and application thereof.

Background

According to the 9 th edition of global diabetes map, (IDF Diabetes Atlas) issued by the international diabetes union, 4.63 million people in the population of 20 to 79 years of age have diabetes mellitus worldwide, most of which are type 2 diabetes mellitus. By 2030 and 2045, it is expected that this number will reach 5.78 hundred million (10.2%) and 7 hundred million (10.9%), respectively. The IDF report indicates: 32% of all diabetics worldwide suffer from cardiovascular disease; over 80% of end-stage renal disease is caused by diabetes or hypertension or both; diabetic foot and lower limb complications affect 4000-6000 ten thousand diabetics.

SGLT-2 inhibitor, chinese name is sodium-glucose cotransporter 2 (sodium-dependent glucose transporters 2, SGLT-2) inhibitor, can inhibit reabsorption of glucose by kidney, and discharge excessive glucose from urine, thereby achieving the purpose of reducing blood sugar. SGLT-2 inhibitors (under the trade name) that have been marketed at present are: faxiga (Dapagliflozin as active ingredient), invoke an (Canagliflozin as active ingredient), gardiifzin (engliflozin as active ingredient), gardiifzin (Empagliflozin as active ingredient), suglat (elgliflozin as active ingredient), and Tofogliflozin formulation product (togliflozin). Both Faxiga, invokana and gardiince of the above mentioned SGLT-2 inhibitors have been approved by the us FDA and EMA for many years and have been demonstrated in combination with diet and exercise to improve glycemic control in adult patients with type II diabetes, reduce the risk of hospitalization for heart failure in adults with type 2 diabetes and cardiovascular disease or multiple cardiovascular risk factors.

The prior art WO03099836A1/CN100534997C discloses methods for the synthesis and purification of dapagliflozin to give dapagliflozin amorphous glassy solids. The glassy solid is easy to soften, form oily matters and absorb moisture to deteriorate, so that active ingredients cannot be uniformly distributed in a product, and the glassy solid cannot be used for preparation production. The preparation, physicochemical properties of the pharmaceutically acceptable crystalline dapagliflozin (S) -propylene glycol hydrate form used in Faxiga, which accompanies desolvation at temperatures between 45 and 100 ℃, are disclosed in WO2008/002824A1/US7919598B2/CN 1014792878. US8513202/CN101573368B discloses that the pharmaceutically acceptable crystalline form used by invoke is canagliflozin hemihydrate, which has a melting point of 97-100 ℃; the preparation of a pharmaceutical crystalline form of tolagliflozin is disclosed in WO2014159151A1, which crystalline form is a hydrate and has a melting point of 71-92 ℃. The crystal form of the active ingredient is suitable for preparation production, and the production process of the preparation composition cannot adopt a powder direct-compression process, otherwise, uneven mixing, sticking and flushing can occur, the content of the active ingredient is not in accordance with the requirements due to electrostatic adsorption, and the like.

The pharmaceutical eutectic is a crystal formed by combining pharmaceutical molecules and eutectic reagents under the action of hydrogen bonds or other noncovalent bonds, and is the focus and the front edge of the current international crystal engineering research.

Research shows that dapagliflozin-L-proline eutectic exists in a polymorphic form, and the preparation process conditions are harsh: the crystallization is carried out for three days at the temperature of 20 ℃ below zero, on one hand, low-temperature control with high energy consumption is required, and meanwhile, three days are required for the eutectic step, so that the whole production period is prolonged. The tolgliflozin-L-proline co-crystal is inferior to a hydrate crystal form with low melting point in crystal form stability, hygroscopicity, static electricity, fluidity, chemical stability and the like. Therefore, the currently marketed products do not adopt the co-crystal of tolagliflozin and L-proline, but adopt the hydrate crystal form with the melting point of 71-92 ℃.

In addition, the ligand or solvent used by the medicinal crystal forms or eutectic crystals of the existing SGLT-2 inhibitor is chiral reagent or chiral ligand, the price is high, and the cost of the finally obtained medicament is high.

The SGLT-2 inhibitor disclosed in the prior art needs to be purified to more than 99% purity by a special purification means before final crystallization, salt formation and eutectic preparation process steps are carried out, and then subsequent crystallization, salt formation and eutectic formation steps are carried out. Especially, for process impurities such as ring-opening impurities, five-membered ring impurities, diastereoisomers, dimers, a large amount of non-specific impurities and the like generated in the preparation process of the SGLT-2 inhibitor, the impurities are difficult to remove by simple solvent recrystallization, and a single purification step is introduced into the process, so that the production period is prolonged, the production cost is obviously improved, and the quality reproducibility and stability between batches in the batch production process are difficult to ensure.

In summary, aiming at the SGLT-2 inhibitor, the development of the pharmaceutical crystal form which is more beneficial to the preparation production, has the advantages of simple crystal form preparation, easy reproduction, low cost and stable and safe quality has great significance for reducing the clinical medication cost and improving the safety, effectiveness and controllability of the products on the market.

Disclosure of Invention

In view of the above, the technical problem to be solved by the invention is to provide an SGLT-2 inhibitor and sarcosine eutectic, and a preparation method and application thereof, and the eutectic of the SGLT-2 inhibitor, which has the advantages of simple process, high repeatability, controllable crystal form and quality, suitability for the mass production of preparation compositions, storage of raw medicines, good stability in the production process and high impurity removal capability, is prepared.

In order to achieve the above purpose, the invention provides an SGLT-2 inhibitor and sarcosine eutectic.

In the present invention, the SGLT-2 inhibitor, which contains classical glucosidic structures, can form co-crystals with sarcosine, preferably, the SGLT-2 inhibitor includes, but is not limited to, dapagliflozin, engagliflozin, canagliflozin, tolagliflozin, iggliflozin, lu Gelie, sogliflozin, canagliflozin, bellgliflozin, elgliflozin, foregliflozin, reggliflozin, taggliflozin, mo Gelie, 2'r,3' r,4's,5's,6 'r) -6- (4-methoxybenzyl) -6' - (hydroxymethyl) -7-methyl-3',4',5',6' -tetrahydropipro [ benzol [ d ] [1,3] dimethyl-4, 2'-pyran ] -3',4',5' -tri, and the specific structural formula is shown in table 1:

TABLE 1 SGLT-2 inhibitor Structure and nomenclature

Preferably, the SGLT-2 inhibitor sarcosine co-crystal is in a substantially crystalline form.

The SGLT-2 inhibitors shown in table 1 above are commonly named after forming co-crystals with sarcosine: dapagliflozin-sarcosine eutectic, engagliflozin-sarcosine eutectic, canagliflozin-sarcosine eutectic, tolagliflozin-sarcosine eutectic, isgliflozin-sarcosine eutectic, lu Gelie mesh-sarcosine eutectic, sogliflozin-sarcosine eutectic, adgliflozin-sarcosine eutectic, begliflozin-sarcosine eutectic, isgliflozin-sarcosine eutectic, hengliflozin-sarcosine eutectic, enggliflozin-sarcosine eutectic, reggliflozin-sarcosine eutectic, mo Gelie mesh-sarcosine eutectic, and the like.

In the SGLT-2 inhibitor-sarcosine co-crystal, the molar ratio of the SGLT-2 inhibitor to the sarcosine is preferably 1: (0.5-5); in some embodiments of the invention, the molar ratio of SGLT-2 inhibitor to sarcosine is preferably 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:0.95, 1:1.0, 1:1.05, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9 or 1:2.0; or a range value having the above ratio as an upper limit or a lower limit; further preferred, the molar ratio of SGLT-2 inhibitor to sarcosine is 1:0.8, 1:0.9, 1:0.95, 1:1.0, 1:1.05, 1:1.1, 1:1.2 or 1:1.3, 1:1.4 or 1:1.5.

The ligand of the SGLT-2 inhibitor and sarcosine eutectic is sarcosine, the sarcosine is a sports nutrition supplement, and the maximum daily dosage of an adult can reach more than 2 g as a food additive or a sports nutrition supplement. Sarcosine is a metabolite of glycine in the human body and is widely distributed in muscles and other tissues of the human body.

The food grade sarcosine has low market price and controllable quality, and the chemical formula is C ₃ H ₇ NO ₂ CAS number 107-97-1, the structural formula is shown as follows:

therefore, the prepared SGLT-2 inhibitor and sarcosine eutectic has higher safety and lower cost.

Preferably, the SGLT-2 inhibitor sarcosine co-crystal has a structure shown in a formula I:

wherein R is ₁ 、R ₂ 、R ₃ 、R ₄ And X is independently selected from the structures shown in Table 2 below:

TABLE 2

In table 2, the broken line indicates the connection position.

Further preferably, the SGLT-2 inhibitor sarcosine co-crystal has a structure represented by formula ii:

wherein R is ₅ 、R ₆ Respectively selected from the structures shown in table 3 below:

TABLE 3 Table 3

The co-crystal of the SGLT-2 inhibitor and the sarcosine can be a solvate or a hydrate of the co-crystal or a solvate hydrate of the co-crystal, the co-crystal of the SGLT-2 and the sarcosine can be characterized by a diffraction angle 2 theta of a characteristic diffraction peak at a specific position in an X-ray powder diffraction (XRPD) pattern, and the model of an XRPD used is as follows: rigakuD/max-RB, test conditions were: the CuK alpha light source, 40kV voltage, 100mA current, slit 1 degree, 0.3mm, and acquisition software is LJ51. Detection was performed using Cu-K alpha radiation.

In the XRPD spectrogram of the eutectic of the SGLT-2 inhibitor and the sarcosine, besides the characteristic diffraction peak of the eutectic part of the SGLT-2 inhibitor, the XRPD spectrogram also has characteristic peaks at the following positions: 10.6 + -0.2 deg., 19.6 + -0.2 deg., 22.1 + -0.2 deg., 33.6 + -0.2 deg..

In some embodiments of the invention, the XRPD pattern of the dapagliflozin-sarcosine co-crystal has characteristic peaks at the following positions: 3.8.+ -. 0.2 °, 10.6.+ -. 0.2 °, 13.7.+ -. 0.2 °, 17.0.+ -. 0.2 °, 18.0.+ -. 0.2 °, 18.6.+ -. 0.2 °, 19.6.+ -. 0.2 °, 20.1.+ -. 0.2 °, 21.4.+ -. 0.2 °, 22.1.+ -. 0.2 °, 23.0.+ -. 0.2 °, 25.4.+ -. 0.2 °, 27.6.+ -. 0.2 °, 33.6.+ -. 0.2 °.

Further preferably, the dapagliflozin-sarcosine co-crystal has characteristic peaks at the following positions using an X-ray powder diffraction (XRPD) pattern expressed in terms of diffraction angle 2θ of Cu-K alpha radiation: 3.77.+ -. 0.2 °, 10.66.+ -. 0.2 °, 11.21.+ -. 0.2 °, 13.67.+ -. 0.2 °, 14.94.+ -. 0.2 °, 16.96.+ -. 0.2 °, 17.98.+ -. 0.2 °, 18.60.+ -. 0.2 °, 19.59.+ -. 0.2 °, 20.10.+ -. 0.2 °, 20.34.+ -. 0.2 °, 21.40.+ -. 0.2 °, 22.10.+ -. 0.2 °, 22.46.+ -. 0.2 °, 22.96.+ -. 0.2 °, 24.87.+ -. 0.2 °, 25.22.+ -. 0.2 °, 25.48.+ -. 0.2 °, 26.23.+ -. 0.2 °, 27.61.+ -. 0.2 °, 28.48.+ -. 0.2 ° and 33.62.+ -. 0.2 °.

In some embodiments of the invention, the XRPD pattern of the canagliflozin-sarcosine co-crystal has characteristic peaks at the following positions: 3.6.+ -. 0.2 °, 7.1.+ -. 0.2 °, 10.6.+ -. 0.2 °, 14.1.+ -. 0.2 °, 16.8.+ -. 0.2 °, 17.3.+ -. 0.2 °, 18.3.+ -. 0.2 °, 18.8.+ -. 0.2 °, 19.6.+ -. 0.2 °, 20.3.+ -. 0.2 °, 21.1.+ -. 0.2 °, 22.1.+ -. 0.2 °, 22.9.+ -. 0.2 °, 25.4.+ -. 0.2 °, 28.2.+ -. 0.2 °, 33.6.+ -. 0.2 °.

Further preferably, the canagliflozin-sarcosine co-crystal uses an X-ray powder diffraction (XRPD) pattern expressed in terms of diffraction angle 2θ of Cu-K alpha radiation, having characteristic peaks at the following positions: 3.63.+ -. 0.2 °, 7.10.+ -. 0.2 °, 10.60.+ -. 0.2 °, 14.08.+ -. 0.2 °, 15.93.+ -. 0.2 °, 16.77.+ -. 0.2 °, 17.34.+ -. 0.2 °, 18.32.+ -. 0.2 °, 18.79.+ -. 0.2 °, 19.60.+ -. 0.2 °, 20.30.+ -. 0.2 °, 20.60.+ -. 0.2 °, 21.11.+ -. 0.2 °, 22.06.+ -. 0.2 °, 22.89.+ -. 0.2 °, 25.41.+ -. 0.2 °, 28.29.+ -. 0.2 ° and 33.46.+ -. 0.2 °.

The invention adopts infrared absorption spectrum to characterize the SGLT-2 inhibitor and sarcosine eutectic structure, for example, KBr tabletting method is adopted to carry out infrared spectrum measurement of SGLT-2 inhibitor and sarcosine eutectic, and the type of an infrared spectrometer is as follows: thermo Nicolet 6700-FT-IR Spectrometer under the following test conditions: KBr pellet method, scanning range is 450-4000cm-1. Characterization is performed by characteristic absorption peaks of infrared spectra.

The infrared spectrum of the SGLT-2 inhibitor sarcosine co-crystal has characteristic absorption peaks at least at the following positions: 3540+ -10 cm ^-1 、2691±5cm ^-1 、2603±5cm ^-1 、2420±5cm ^-1 。

In some embodiments of the invention, the infrared spectrum of the dapagliflozin-sarcosine co-crystal has characteristic absorption peaks at the following positions: 3543.67 + -10 cm ^-1 、3161.94±10cm ^-1 、2690.95±5cm ^-1 、2603.78±5cm ^-1 、2419.39±5cm ^-1 、2360.665cm ^-1 、1599.61±5cm ^-1 、1510.92±5cm ^-1 、1291.19±5cm ^-1 、1045.97cm ^-1 。

In some embodiments of the invention, the infrared spectrum of the canagliflozin-sarcosine co-crystal has characteristic absorption peaks at least at the following positions: 3543.82 + -10 cm ^-1 、3153.82±10cm ^-1 、2690.68±5cm ^-1 、2603.06±5cm ^-1 、2419.53±5cm ^-1 、1596.33±5cm ^-1 、1507.48±5cm ^-1 、1086.11±5cm ^-1 、1062.19±5cm ^-1 、829.61±5cm ^-1 。

The SGLT-2 inhibitor-sarcosine eutectic is characterized by a differential scanning calorimetry method, and the model of an instrument is as follows: SII-DSC6220, analytical method parameters: temperature range: scanning rate at 30-250 deg.c: 10 ℃/min, shielding gas: nitrogen, 50 ml/min. The obtained Differential Scanning Calorimeter (DSC) spectrum has a distinct endothermic peak at 100℃or more, preferably 120℃or more.

According to Differential Scanning Calorimetric (DSC) spectrum data, the melting point of the dapagliflozin-sarcosine eutectic crystal provided by the invention is about 149.0 ℃ which is higher than the melting point of the dapagliflozin (S) -propylene glycol monohydrate which is already marketed, and the proper melting point ensures that the preparation is not easy to melt and agglomerate in the processes of granulating and tabletting tablets, so that sticking or uniformity is not qualified.

The SGLT-2 inhibitor and sarcosine eutectic provided by the invention is detected by adopting a thermogravimetric analysis method, and the model of an instrument of the thermogravimetric analyzer is as follows: SII-TG/DTA6200, analytical method parameters: temperature range: scanning rate at 30-350 deg.c: 10 ℃/min, shielding gas: nitrogen, 200 ml/min. No significant weight loss in thermogravimetric analysis (TGA) spectra before 120 ℃; has a wider endothermic peak at 190-230 ℃ in the TGA-DTA spectrogram.

The invention also detects the SGLT-2 inhibitor and sarcosine eutectic structure through nuclear magnetic resonance hydrogen spectrum. The instrument model used was: bruker avance600, resonance frequency: 600MHz, using solvent: deuterated methanol. As a result, it was found that in the hydrogen nuclear magnetic resonance spectrum (HNMR), in addition to the formants having the structure of SGLT-2 inhibitor, there was-CH in the range of 2.4 to 3.2ppm in chemical shift ₃ Has a formant of-CH in the range of 3.0-4.0ppm ₂ -peak, -CH ₂ Peaks may or may not coincide with other formants on the SGLT-2 structure, indicating the presence of sarcosine results.

The invention provides a preparation method of the SGLT-2 inhibitor and sarcosine eutectic, which comprises the following steps:

mixing the SGLT-2 inhibitor solution and the sarcosine solution, standing for crystallization or cooling for crystallization, and carrying out solid-liquid separation to obtain the SGLT-2 inhibitor-sarcosine eutectic.

In the present invention, the source of the SGLT-2 inhibitor is not particularly limited, and it may be generally commercially available or prepared according to a method well known to those skilled in the art, and it may be pure or crude or intermediate.

Preferably, the solvent in the SGLT-2 inhibitor solution is selected from different single solvents or mixed solvents of C1-C10 alcohols, C3-C10 ketones, ethers and nitriles. Further preferably, the solvent is one or more of ethanol, acetone, tetrahydrofuran and acetonitrile; ethanol is more preferred.

The solvent of the sarcosine solution is preferably water.

The molar ratio of the SGLT-2 inhibitor to sarcosine is preferably 1: (0.5 to 5.0), more preferably 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:0.95, 1:1.0, 1:1.05, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9 or 1:2.0.

in the process of preparing the co-crystal, the SGLT-2 inhibitor and the sarcosine are added in a strict molar ratio according to the two in the co-crystal, so that the SGLT-2 inhibitor and the sarcosine just form the co-crystal, and no excessive SGLT-2 inhibitor or sarcosine exists in the system.

In the present invention, after the SGLT-2 inhibitor solution and the sarcosine solution are mixed, if the system is not clarified, the system may be clarified by heating.

In the present invention, the temperature of the standing crystallization or the cooling crystallization is preferably-20 ℃ to 40 ℃, and in some embodiments, the crystallization temperature is preferably: -15-35 ℃, 10-30 ℃, 5-30 ℃, 0-30 ℃, 5-30 ℃, 10-30 ℃, 15-30 ℃ or 20-30 ℃.

In the present invention, the time for standing crystallization or cooling crystallization is preferably 4 to 48 hours, preferably 4 to 24 hours, 4 to 16 hours, 4 to 12 hours, more preferably 8 to 12 hours.

In the present invention, after the SGLT-2 inhibitor/sarcosine co-crystal is obtained, it is preferably subjected to a drying treatment at a temperature of preferably 20 to 80 ℃, more preferably 30 to 80 ℃.

The SGLT-2 inhibitor sarcosine co-crystal obtained may be a solvate of the co-crystal, a solvate hydrate of the co-crystal, or a hydrate of the co-crystal.

The preparation method of the SGLT-2 inhibitor and sarcosine eutectic provided by the invention has the advantages of simple process, easiness in realization and short period, and the obtained crystal form is stable and is not influenced by crystallization conditions or storage conditions or environment, so that the phenomenon of crystal transformation or mixed crystal is generated.

The SGLT-2 inhibitor-sarcosine eutectic prepared by the invention has no more than 0.1 percent of single impurity, and the total amount of the impurities is less than 0.5 percent.

In the invention, the crude product (or called intermediate) of the SGLT-2 inhibitor is mixed with sarcosine, stirred at room temperature and subjected to solid-liquid separation to obtain the sarcosine compound of the SGLT-2 inhibitor, which has remarkable purification effect on the crude product of the SGLT-2 inhibitor, and the purity of the SGLT-2 inhibitor is not lower than 99 percent after purification.

Because most of the chemical structures of SGLT-2 inhibitors have classical glucose glucoside structures or glucoside structure derivatives, corresponding isomer impurities, ring-opening impurities, five-membered ring impurities or dimer impurities are inevitably generated in the preparation process, and therefore the removal capacity of salifying, solvating or hydrating specific process impurities, nonspecific process impurities and process byproducts has great influence on the preparation yield, production period and production cost of active ingredients.

For example, diastereoisomeric impurities produced during the preparation process have the following reaction equation:

or the following impurity 2 (five membered ring impurity) is also generated in the above SGLT-2 inhibitor preparation process:

The generation mechanism of the five-membered ring impurity is as follows:

or the following impurity 3 (ring-opening impurity) is also generated in the above SGLT-2 inhibitor preparation process:

the generation process of ring-opening impurities is as follows:

or dimer impurity 4 (dimer impurity) is produced by intermolecular polymerization reaction due to severe reaction conditions in the above SGLT-2 inhibitor production process:

the chemical structural formula of the dimer impurity is as follows:

the production process of the dimer impurity is as follows:

wherein X is halogen (bromine Br or iodine I); m is magnesium, mg; p is a protecting group such as tetramethylsilane TMS.

R ₁ ～R ₅ The ranges of (2) are the same and are not described in detail herein.

In addition to the process impurities which are generated due to the specificity (diastereoisomeric impurities, five-membered ring impurities, ring-opening impurities and dimer impurities) related to the classical glucoside structure in the SGLT-2 inhibitor structure, a large amount of non-specific impurities are generated in the process, and the specific impurities and the non-specific impurities are difficult to be removed below the pharmaceutically acceptable impurity limit in the SGLT-2 inhibitor intermediate or crude product at one time through simple extraction and solvent recrystallization.

Surprisingly, by using the method of the invention, the sarcosine compound is formed by the SGLT-2 inhibitor crude product and sarcosine, more than 95% of impurities such as process impurities, intermediates in the last step or material residues and the like contained in the SGLT-2 inhibitor crude product can be removed, the impurities have similar polarity to the target API and similar structure, the purification purpose is difficult to achieve by simple liquid-liquid extraction or solvent recrystallization, the sarcosine compound is formed by the SGLT-2 inhibitor crude product and the sarcosine, the purity of the SGLT-2 inhibitor crude product can be increased from 78% to more than 99% at one time, and the number of the impurities and the total impurity detection amount are obviously reduced. Or the sarcosine compound of the high-purity SGLT-2 inhibitor is subjected to simple dissociation to obtain the higher-purity free SGLT-2 inhibitor, and then the higher-purity free SGLT-2 inhibitor is directly used as a target component or is used for preparing other medicinal crystal forms or medicinal co-crystals.

Specifically, the invention provides a purification method of an SGLT-2 inhibitor, which comprises the following steps:

mixing the crude product of the SGLT-2 inhibitor with sarcosine, stirring at room temperature, and carrying out solid-liquid separation to obtain the sarcosine compound of the SGLT-2 inhibitor.

Preferably, the crude SGLT-2 inhibitor is dissolved in a solvent, preferably a different single solvent of a C1-C10 alcohol, a C3-C10 ketone, an ether, a nitrile, or a mixed solvent of these solvents and water or a mixed solvent of the above solvents. Further preferably, the solvent is one or more of ethanol, acetone, tetrahydrofuran and acetonitrile, and can also be one or more of ethanol, acetone, tetrahydrofuran and acetonitrile mixed with water; ethanol is more preferred.

Preferably, the sarcosine is dissolved in a solvent, preferably water.

The molar ratio of the crude SGLT-2 inhibitor to sarcosine is preferably 1: (0.5 to 5.0), in some embodiments, the molar ratio of crude SGLT-2 inhibitor to sarcosine is preferably 1:0.7, 1:0.8, 1:0.9, 1:0.95, 1:1.0, 1:1.05, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2.0 or 1:3.0; more preferred crude SGLT-2 inhibitor is present in a molar ratio to sarcosine of 1:1.0, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2.0 and 1:3.0.

In the present invention, the above-mentioned sarcosine is excessively added during the purification process, and the resultant sarcosine compound of the SGLT-2 inhibitor contains a part of the co-crystal formed by the SGLT-2 inhibitor and the sarcosine, and a part of the excessive sarcosine.

In the present invention, the solid-liquid separation method is preferably standing crystallization or cooling crystallization.

In the present invention, the temperature of the standing crystallization or the cooling crystallization is preferably-20℃to 40℃and more preferably-10℃to 40℃and 0-40℃and 10-40℃and 15-40℃and 20-35℃or 20℃to 30 ℃.

Preferably, the stirring time is 6 to 16 hours, more preferably 6 to 14 hours, 7 to 13 hours, or 8 to 12 hours.

In the present invention, the SGLT-2 inhibitor and the sarcosine compound are obtained, and a solid is obtained by solid-liquid separation, and preferably, the solid is subjected to drying treatment, and the drying temperature is preferably 20 to 80 ℃, and more preferably 30 to 80 ℃.

Preferably, the method further comprises the steps of:

and (3) dissociating the sarcosine compound of the SGLT-2 inhibitor to obtain a pure product of the SGLT-2 inhibitor in a free state.

Preferably, the HPLC normalized purity of the free pure product of the SGLT-2 inhibitor is not less than 99%.

The final pharmaceutical crystalline form of the SGLT-2 inhibitor is then prepared by recrystallization or eutectic means.

Preferably, the final pharmaceutically acceptable crystal form of the SGLT-2 inhibitor is selected from pure, solvate, hydrate, solvate hydrate, co-crystal or double salt of the SGLT-2 inhibitor.

The process or source of the crude SGLT-2 inhibitor is not particularly limited, and may be one prepared according to methods known to those skilled in the art, for example, the methods described in WO03099836A1 and CN 100534997C. The SGLT-2 inhibitor of the following structural formula 1 should be contained in the crude SGLT-2 inhibitor or intermediate not less than 70%, preferably not less than 75%.

Wherein R1, R2, R3, R4 and X are as shown in Table 2.

The method of dissociation in the present invention is not particularly limited, and may be a method well known to those skilled in the art.

In some embodiments of the invention, dissociation may be performed by liquid-liquid extraction, and the organic phase is collected and concentrated to obtain pure SGLT-2 inhibitor in its free form.

Experimental results show that the purification method provided by the invention has obvious removal effects on non-specific impurities, diastereoisomeric impurities, five-membered ring impurities, ring-opening impurities and dimer impurities. The number of impurities is reduced by more than 90%, and the total amount of the detected impurities, the single impurity detection amount and the like are removed by more than 95%. The number of impurities in the sarcosine compound of the SGLT-2 inhibitor or the free body of the SGLT-2 inhibitor obtained after purification is less than 6, and the total impurity content is less than 1.0%.

The purification method provided by the invention can be used for an intermediate in the preparation process of the SGLT-2 inhibitor, and can also be used for a crude product before salifying, eutectic forming and crystallization, and preferably an intermediate or a crude product of the intermediate of the SGLT-2 inhibitor, the chemical structure of which is the same as that of a target SGLT-2 inhibitor.

Preferably, after the pure free form product of the SGLT-2 inhibitor is obtained, the pure free form product of the SGLT-2 inhibitor can be used as a raw material to directly prepare a final medicinal crystal form of the SGLT-2 inhibitor;

or the pure free SGLT-2 inhibitor is used as a raw material, and the final medicinal crystal form of the SGLT-2 inhibitor is prepared through recrystallization or eutectic mode.

In the invention, the final medicinal crystal form of the SGLT-2 inhibitor is preferably a pure product, solvate, hydrate, solvate hydrate, co-crystal or double salt of the SGLT-2 inhibitor.

The invention provides a pharmaceutical composition, which comprises the SGLT-2 inhibitor and sarcosine eutectic, or the SGLT-2 inhibitor and sarcosine eutectic prepared by the preparation method, or the free pure product of the SGLT-2 inhibitor obtained by the purification method, or the final medicinal crystal form of the SGLT-2 inhibitor prepared by recrystallization or eutectic mode, such as the pure product, solvate, hydrate, solvate hydrate, eutectic or double salt of the SGLT-2 inhibitor, and pharmaceutically acceptable carriers, excipients, diluents, auxiliary agents, vehicles or the combination thereof.

When the active ingredient is administered in low doses, the particles of the active ingredient may be reduced to a suitable size by some physical means, such as grinding, sieving, in order to ensure a uniform distribution of the active ingredient. The SGLT-2 inhibitor-sarcosine eutectic provided by the invention has higher stability, and is favorable for being uniformly distributed in a medicinal composition product as an active ingredient. The composition has a higher melting point, does not melt or agglomerate or electrostatic adsorption and the like due to heat in the grinding or milling process, and the crystal form is not changed in the whole grinding or milling process, so that the finally obtained pharmaceutical composition has uniform content of active ingredients, accords with the marking amount and has higher repeatability.

The invention provides an SGLT-2 inhibitor and sarcosine co-crystal or an SGLT-2 inhibitor and sarcosine co-crystal prepared by the preparation method, or an SGLT-2 inhibitor free pure product obtained by the purification method, or a final medicinal crystal form of the SGLT-2 inhibitor prepared by recrystallization or eutectic mode, such as an SGLT-2 inhibitor pure product, a solvate, a hydrate, a solvate hydrate, a co-crystal or a double salt, or the like, or the pharmaceutical composition, and application thereof in preparing medicines for preventing, treating or relieving diabetes and complications thereof, and treating or preventing or relieving cardiovascular and cerebrovascular diseases and non-diabetes-induced nephropathy.

The invention provides an SGLT-2 inhibitor and sarcosine eutectic or an SGLT-2 inhibitor and sarcosine eutectic prepared by the preparation method, or an SGLT-2 inhibitor free pure product obtained by the purification method, or a final medicinal crystal form of the SGLT-2 inhibitor, such as an SGLT-2 inhibitor pure product, a solvate, a hydrate, a solvate hydrate, a eutectic or a double salt, or the like, prepared by recrystallization or a eutectic mode, or application of the pharmaceutical composition in preparation of a medicament for reducing blood pressure.

Preferably, the diabetes and its complications include, but are not limited to:

primary hypertension, type 2 diabetes mellitus with hypertension, kidney disease with type 2 diabetes mellitus, kidney disease with hypertension and diabetes mellitus, kidney disease, type 1 diabetes, kidney disease with type 1 diabetes mellitus, liver fibrosis, insulin resistance, hyperglycemia, hyperinsulinemia, elevated blood levels of fatty acids or glycerol, hyperlipidemia, dyslipidemia, obesity.

The SGLT-2 inhibitor and sarcosine co-crystal, or the free pure SGLT-2 inhibitor obtained by the purification method, or the final medicinal crystal forms of the SGLT-2 inhibitor, such as the pure SGLT-2 inhibitor, solvate, hydrate, solvate hydrate, co-crystal or double salt, or the like, are prepared by recrystallization or co-crystallization, or the pharmaceutical composition can be used singly or in combination with other medicines when being applied to preparing medicines for preventing, treating or relieving diabetes and complications thereof, cardiovascular and cerebrovascular diseases, or medicines for reducing blood pressure or medicines for treating renal diseases caused by non-diabetes.

The invention provides a method for preventing, treating or relieving diabetes and complications thereof, which comprises the steps of contacting the SGLT-2 inhibitor and sarcosine eutectic or the SGLT-2 inhibitor and sarcosine eutectic prepared by the preparation method, or the free pure SGLT-2 inhibitor obtained by the purification method, or the final medicinal crystal form of the SGLT-2 inhibitor, such as the pure SGLT-2 inhibitor, solvate, hydrate, solvate hydrate, eutectic or double salt, or the like, prepared by recrystallization or eutectic mode, or the pharmaceutical composition with biological samples.

The invention provides a method for reducing blood pressure, which comprises the steps of contacting the SGLT-2 inhibitor and sarcosine eutectic or the SGLT-2 inhibitor and sarcosine eutectic prepared by the preparation method, or the free pure product of the SGLT-2 inhibitor obtained by the purification method, or the final medicinal crystal form of the SGLT-2 inhibitor, such as the pure product, solvate, hydrate, solvate hydrate, eutectic or double salt of the SGLT-2 inhibitor, or the pharmaceutical composition with biological samples.

The invention provides a method for treating non-diabetes-induced nephropathy, which comprises the steps of contacting the SGLT-2 inhibitor and sarcosine eutectic or the SGLT-2 inhibitor and sarcosine eutectic prepared by the preparation method, or the free pure SGLT-2 inhibitor obtained by the purification method, or the final medicinal crystal form of the SGLT-2 inhibitor, such as the pure SGLT-2 inhibitor, solvate, hydrate, solvate hydrate, eutectic or double salt, or the like, or the medicinal composition with biological samples.

Compared with the prior art, the invention provides an SGLT-2 inhibitor and sarcosine eutectic. Has the following beneficial effects:

1. the sarcosine is used as a ligand, so that the method has higher safety and lower cost;

2. the pharmaceutical co-crystal has higher stability, and the crystal form is not easy to change in the production process or the storage process of the preparation composition; the temperature in the storage or preparation process is high or the preparation process can not melt when meeting heat, the sticking, agglomeration or static electricity can not occur, and the mixing uniformity is better;

3. as an active ingredient, the active ingredient is uniformly distributed in the prepared pharmaceutical composition, so that the pharmaceutical composition has better in vivo release, absorption and drug effect and small batch-to-batch difference;

4. the sarcosine co-crystal of the SGLT-2 inhibitor does not change the solubility of a medicine used as a bulk medicine of an oral solid preparation in physiological media with different pH values, which is very important for ensuring the absorption of the oral solid preparation;

5. the product has high physical stability and chemical stability, and is more favorable for storage and transportation;

6. the preparation process is simple, the repeatability is high, the crystallization time is short, the process condition requirement is low, and the economic benefit is higher;

7. in the preparation process, unsafe solvents are not used;

8. In the preparation process of the SGLT-2 inhibitor, the final intermediate or crude product of the SGLT-2 inhibitor can be purified and dissociated through a sarcosine compound to obtain an SGLT-2 inhibitor free body with the purity of more than 99 percent, and then the SGLT-2 inhibitor free body is subjected to subsequent recrystallization, eutectic or eutectic with sarcosine according to the requirement of a target API, so that the complex impurity removal step is not needed.

Drawings

FIG. 1 is an X-ray powder diffraction pattern (XRPD) and single-crystal diffraction pattern of dapagliflozin-sarcosine co-crystal prepared in example 1;

FIG. 2 is an infrared spectrum analysis chart (IR) of dapagliflozin-sarcosine eutectic prepared in example 1;

FIG. 3 is a Differential Scanning Calorimeter (DSC) of dapagliflozin-sarcosine co-crystal prepared in example 1;

FIG. 4 is a thermogravimetric analysis (TGA) of dapagliflozin-sarcosine co-crystal prepared in example 1;

FIG. 5 is a nuclear magnetic resonance hydrogen spectrum (H-NMR) of dapagliflozin-sarcosine eutectic prepared in example 1;

FIG. 6 is an X-ray powder diffraction pattern (XRPD) of the co-crystal of canagliflozin and sarcosine produced in example 12;

FIG. 7 is an infrared spectrum analysis chart (IR) of the co-crystal of canagliflozin and sarcosine prepared in example 12;

FIG. 8 is a Differential Scanning Calorimeter (DSC) of a co-crystal of canagliflozin and sarcosine prepared in example 12;

FIG. 9 is a thermogravimetric analysis (TGA) of the canagliflozin-sarcosine co-crystal prepared in example 12;

FIG. 10 is a nuclear magnetic resonance hydrogen spectrum (H-NMR) of a canagliflozin-sarcosine co-crystal prepared in example 12;

FIG. 11 is an ellipsometric view of the molecular structure of the single crystal structure of dapagliflozin-sarcosine eutectic.

Detailed Description

The raw material of sarcosine used in the present invention is commercially available.

The dapagliflozin starting material used in the present invention is commercially available or can be prepared according to a known method, for example, the method described in patent document CN 100534997C.

The solvent used in the present invention is not particularly limited, and commercially available conventional solvents can be used.

The instrument and the method for collecting data are as follows:

x-ray powder diffraction (XRPD) data were collected using the instrument model number: rigakuD/max-RB, test conditions were: the CuK alpha light source, 40kV voltage, 100mA current, slit 1 degree, 0.3mm, and acquisition software is LJ51.

Instrument model of X-single crystal diffraction: the Gemini E single crystal diffractometer has crystal analysis software of Shellx 97; cuKa as the X-ray source.

Infrared spectroscopic analysis (IR) data were collected using the instrument model: thermo Nicolet 6700-FT-IR Spectrometer under the following test conditions: KBr tabletting method, scanning range is 450-4000cm ^-1 。

Differential Scanning Calorimeter (DSC) data were collected using the instrument model: SII-DSC6220, analytical method parameters: temperature range: scanning rate at 30-250 deg.c: 10 ℃/min, shielding gas: nitrogen, 50 ml/min.

Thermogravimetric analysis (TGA) data was collected using the instrument model: SII-TG/DTA6200, analytical method parameters: temperature range: scanning rate at 30-350 deg.c: 10 ℃/min, shielding gas: nitrogen, 200 ml/min.

Nuclear magnetic resonance hydrogen spectrum (HNMR) data were collected using the instrument model: bruker avance600, resonance frequency: 600MHz, using solvent: deuterated methanol.

The liquid phase test conditions related to the invention are: the chromatographic column is Kromasil KR100-5-C18, 4.6X1250 mm; mobile phase a:0.1% phosphoric acid; mobile phase B: acetonitrile; detection wavelength: 220nm; flow rate: 0.8mL/min; sample injection amount: 20. Mu.L; column temperature: 35 ℃; the liquid phase conditions are shown in table 4 below:

table 4 liquid phase test conditions

t(min)	0	25	30	35	40	45	45.1	60
B(％)	30	45	50	70	70	90	30	30

It should be noted that the numerical values or numerical endpoints of the present invention are included in the scope of the present invention, and the meaning and meaning of the numerical values or endpoints are not limited to the numerical values themselves, and those skilled in the art will appreciate that they include those allowable error ranges such as experimental error, measurement error, statistical error, random error, etc., which are well accepted in the art, and all such error ranges are included in the scope of the present invention.

For further explanation of the present invention, the SGLT-2 inhibitor-sarcosine co-crystal, the preparation method and application thereof provided by the present invention are described in detail with reference to the following examples, which do not limit the scope of the present invention in any way.

In the following examples, all reagents, canagliflozin, sarcosine, etc., are generally commercially available, unless otherwise indicated.

Dapagliflozin used in the following examples was prepared according to patent document WO03099836 A1.

EXAMPLE 1 preparation of dapagliflozin-sarcosine Co-crystals

500mL of absolute ethanol is added into a 1000mL three-necked flask, stirring is started, 100g of dapagliflozin (HPLC normalized purity is more than or equal to 95%) prepared according to the method provided in patent document WO03099836A1 is added, and stirring is carried out until the system is dissolved. Another 100mL three-necked flask was taken, 30mL of purified water was added, stirring was started, 21.80g of sarcosine was added, and after stirring until the system was clear, the solution was added to the above dapagliflozin ethanol solution. The mixture is stirred and crystallized for 5 hours at the temperature of 20 ℃ to 30 ℃, filtered, and the collected solid is dried in vacuum for 6 hours at the temperature of 30 ℃ to 40 ℃ to obtain 113g white solid, wherein the normalized purity of HPLC is 99.93 percent, and the impurity with the normalized content exceeding 0.1 percent is not generated.

The dapagliflozin-sarcosine co-crystal was confirmed by X-ray powder diffraction pattern (XRPD), infrared spectrum analysis pattern (IR), differential Scanning Calorimeter (DSC), thermogram (TGA), nuclear magnetic resonance hydrogen spectrum (H-NMR), and detailed in fig. 1 to 5.

¹ HNMR(600MHz,MeOD)δ7.343-7.329(d，1H，J＝8.4Hz)，7.315-7.312(d，1H，J＝1.8Hz)7.276-7.259(dd，1H，J＝8.4，1.8Hz)，7.091-7.077(d，2H，J＝8.4Hz)，6.795-6.781(d，2H，J＝8.4Hz)，4.089-4.073(d，1H，J＝9.6Hz)，4.055-3.977(q，2H，J＝15.0Hz)，3.994-3.959(q，2H，J＝7.2Hz)，3.876-3.854(dd，1H，J＝12.0，1.8Hz)，3.697-3.668(dd，1H，J＝12.0，5.4Hz)，3.467(s，2H)3.461-3.431(m，1H)，3.409-3.370(m，2H)，3.283-3.268(m，1H)，2.665(s，3H)，1.360-1.337(t，3H，J＝7.2Hz)。

According to ¹ HNMR spectrum data shows that the molar ratio of dapagliflozin to sarcosine is 1:1.

The dapagliflozin-sarcosine co-crystal, using an X-ray powder diffraction (XRPD) pattern of Cu-K alpha radiation expressed in terms of diffraction angle 2θ, has characteristic peaks and their relative intensities as shown in table 5 below, with diffraction angle 2θ error of ± 0.2 °:

TABLE 5 dapagliflozin sarcosine eutectic XRPD characteristic peaks and relative intensities thereof

No.	Diffraction angle 2 theta	D	I/Io	No.	Diffraction angle 2 theta	D	I/Io
1	3.765	23.4510	72.3	10	20.343	4.3618	57.3
2	10.656	8.2953	40.8	11	21.404	4.1479	32.2
3	11.210	7.8862	16.4	12	22.096	4.0196	16.7

4	13.674	6.4705	51.0	12	22.962	3.8699	100
5	14.939	5.9253	17.7	13	25.482	3.4927	29.0
6	16.956	5.2247	44.8	14	27.606	3.2285	60.6
7	17.977	4.9301	89.6	15	30.435	2.9346	15.3
8	18.602	4.7658	51.6	16	33.617	2.6637	12.6-
9	19.588	4.5283	95.2	--	--	--	--

The dapagliflozin-sarcosine eutectic has an infrared absorption (IR) spectrum with absorption peaks at the following positions measured by KBr tabletting: 3543.67 + -10 cm ^-1 、3161.94±10cm ^-1 、2888.82±5cm ^-1 、2690.95±5cm ^-1 、2603.78±5cm ^-1 、2419.39±5cm ^-1 、2360.66±5cm ^-1 、1599.61±5cm ^-1 、1510.92±5cm ^-1 、1291.19±5cm ^-1 、1045.97±5cm ^-1 、812.44±5cm ^-1 cm ^-1 。

The dapagliflozin-sarcosine eutectic has a Differential Scanning Calorimeter (DSC) spectrum, has an endothermic peak in the range of 140.0-155.0 ℃, the peak value is 149.0 ℃, and the melting temperature is the melting temperature.

The thermogravimetric analysis (TGA) profile of dapagliflozin-sarcosine co-crystal, without significant weight loss before 150 ℃; the TGA-DTA diagram has a wider endothermic peak in the range of 190-230 ℃. The water content was shown from thermogravimetric analysis (TGA) spectra, and moisture meter (instrument model: metrohm 915-KF): 0.10% shows that the dapagliflozin-sarcosine eutectic crystal is anhydrous and exists in a non-solvate form.

Example 2: preparation of dapagliflozin-sarcosine eutectic

50mL of absolute ethanol is added into a 100mL three-necked flask, stirring is started, 10g of dapagliflozin (HPLC normalized purity is more than or equal to 95%) prepared according to the method provided in patent document WO03099836A1 is added, and stirring is carried out until the system is dissolved. Another 10mL single-mouth bottle was taken, 2mL of purified water was added, stirring was started, 1.10g of sarcosine was added, and after stirring until the system was clear, this solution was added to the ethanol solution of dapagliflozin described above. The mixture is stirred and crystallized for 5 hours at the temperature of 10 ℃ to 20 ℃, filtered, and the collected solid is dried in vacuum for 4 hours at the temperature of 40 ℃ to 50 ℃ to obtain 4.8g of white solid reaching the gliflozin-sarcosine eutectic, the HPLC normalized purity is 99.89%, and no impurity with the normalized content exceeding 0.1 percent is produced.

Example 3: preparation of dapagliflozin-sarcosine eutectic

50mL of absolute ethanol is added into a 100mL three-necked flask, stirring is started, 10g of dapagliflozin (HPLC normalized purity is more than or equal to 95%) prepared according to the method provided in patent document WO03099836A1 is added, and stirring is carried out until the system is dissolved. Another 10mL single-port bottle was taken, 5.0mL of purified water was added, stirring was started, 3.27g of sarcosine was added, and after stirring until the system was clear, the solution was added to the ethanol solution of dapagliflozin. The mixture is stirred and crystallized for 5 hours at the temperature of-10 ℃ to 10 ℃, filtered, and the collected solid is dried in vacuum for 4 hours at the temperature of 50 ℃ to 60 ℃ to obtain 10.85g of white solid reaching the gliflozin-sarcosine eutectic, the HPLC normalized purity is 99.87%, and no impurity with the normalized content exceeding 0.1 percent is produced.

Example 4: preparation of dapagliflozin-sarcosine eutectic

50mL of acetonitrile was added to a 100mL three-necked flask, stirring was started, 10g of dapagliflozin (HPLC normalized purity: 95%) prepared by the method provided in patent document WO03099836A1 was added, and the mixture was stirred to a system supernatant. Another 10mL single-port bottle was taken, 3.0mL of purified water was added, stirring was started, 2.18g of sarcosine was added, and after stirring until the system was clear, this solution was added to the acetonitrile solution of dapagliflozin described above. The mixture is stirred and crystallized for 5 hours at the temperature of 30-40 ℃, filtered, and the collected solid is dried in vacuum for 4 hours at the temperature of 70-80 ℃ to obtain 9.70g of white solid reaching the gliflozin-sarcosine eutectic, the HPLC normalized purity is 99.90%, and no impurity with the normalized content exceeding 0.1 percent is produced.

Example 5: preparation of dapagliflozin-sarcosine eutectic

50mL of tetrahydrofuran was added to a 100mL three-necked flask, stirring was started, 10g of dapagliflozin (HPLC normalized purity: 95%) prepared by the method described in patent document WO03099836A1 was added, and the mixture was stirred to a system supernatant. Another 10mL single-port bottle was taken, 3.0mL of purified water was added, stirring was started, 2.20g of sarcosine was added, and after stirring until the system was clear, this solution was added to the above dapagliflozin in tetrahydrofuran. The mixture is stirred and crystallized for 5 hours at the temperature of 20 ℃ to 30 ℃, filtered, and the collected solid is dried in vacuum for 4 hours at the temperature of 60 ℃ to 70 ℃ to obtain 9.15g of white solid reaching the gliflozin sarcosine eutectic, the HPLC normalized purity is 99.73%, and no impurity with the normalized content exceeding 0.1 percent is produced.

Example 6: preparation of dapagliflozin-sarcosine eutectic

50mL of acetone was added to a 100mL three-necked flask, stirring was started, 10g of dapagliflozin (HPLC normalized purity: 95%) prepared by the method described in patent document WO03099836A1 was added, and the mixture was stirred until the system was clear. Another 10mL single-port bottle was taken, 3.0mL of purified water was added, stirring was started, 2.10g of sarcosine was added, and after stirring until the system was clear, this solution was added to the acetone solution of dapagliflozin. The mixture is stirred and crystallized for 5 hours at the temperature of 20-30 ℃, filtered, and the collected solid is dried in vacuum for 6 hours at the temperature of 20-30 ℃ to obtain 10.13g of white solid reaching the gliflozin-sarcosine eutectic, the HPLC normalized purity is 99.82%, and no impurity with the normalized content exceeding 0.1 percent is produced.

Example 7: preparation of dapagliflozin-sarcosine eutectic

To a 100mL three-necked flask, 25mL of acetone and 25mL of absolute ethanol were added, stirring was started, 10g of dapagliflozin (HPLC normalized purity: 95% or more) prepared according to the method provided in patent document WO03099836A1 was added, and the mixture was stirred until the system was clear. Another 10mL single-port bottle was taken, 3.0mL of purified water was added, stirring was started, 2.15g of sarcosine was added, and after stirring until the system was clear, this solution was added to the above dapagliflozin in acetone and ethanol solution. The mixture is stirred and crystallized for 5 hours at the temperature of 20-30 ℃, filtered, and the collected solid is dried in vacuum for 6 hours at the temperature of 30-40 ℃ to obtain 10.89g of white solid reaching the gliflozin-sarcosine eutectic, the HPLC normalized purity is 99.82%, and no impurity with the normalized content exceeding 0.1 percent is produced.

Example 8: purification of dapagliflozin crude product

150mL of absolute ethanol was added to a 250mL three-necked flask, stirring was started, 35g of dapagliflozin crude product prepared by the method provided in patent document WO03099836A1 (HPLC normalized purity: 78.04%) was added, and the mixture was stirred to a system supernatant. Another 50mL single-mouth bottle was taken, 10mL of purified water was added, stirring was started, 9.00g of sarcosine was added, and after stirring until the system was clear, the solution was added to the above dapagliflozin solution in absolute ethanol. The mixture is stirred and crystallized for 5 hours at the temperature of 20 to 30 ℃, filtered, and the collected solid is dried in vacuum for 6 hours at the temperature of 30 to 40 ℃ to obtain 25.4g of dapagliflozin sarcosine compound which is white solid with the HPLC normalized purity of 99.37 percent.

To a 250mL three-necked flask, 50mL of purified water and 150mL of methyl tert-butyl ether were added, 25g of the above-obtained dapagliflozin sarcosine compound was added, the organic phase was collected by stirring, the organic phase was washed 3 times with 50mL of purified water, the solvent was distilled off from the organic phase under reduced pressure, and vacuum-dried at 30℃to 40℃for 6 hours, to obtain 19.5g of dapagliflozin as a white solid (HPLC normalized purity 99.98%).

Comparative example 1:

600mL of methyl tert-butyl ether was added to a 3000mL three-necked flask, stirring was started, 35g of dapagliflozin crude product prepared by the method provided in patent document WO03099836A1 (HPLC normalized purity: 78.04%) was added, stirred to a system supernatant, 6.51g of (S) -1, 2-propanediol, 1.54g of purified water, 0.35g of dapagliflozin (S) -propanediol monohydrate seed crystal were further added by the method of example 6 for preparing dapagliflozin (S) -propanediol monohydrate according to patent document WO2008002824A1, crystallization was carried out at 20℃to 30℃for 5 hours with stirring, 1200mL of cyclohexane was added, filtration was carried out, and the collected solid was dried in vacuo at 20℃to 25℃for 4 hours to obtain 25g of off-white solid (HPLC normalized purity: 93.59%).

To a 250mL three-necked flask, 50mL of purified water and 150mL of methyl t-butyl ether were added, 20g of dapagliflozin (S) -propylene glycol monohydrate obtained above was added, the organic phase was separated by stirring, the organic phase was washed 3 times with 50mL of purified water, and the solvent was distilled off under reduced pressure to obtain 18g of dapagliflozin as a pale yellow foamy solid (HPLC normalized purity: 95.78%).

Example 9: purification of dapagliflozin intermediates

150mL of absolute ethanol was added to a 250mL three-necked flask, stirring was started, and 30g of crude dapagliflozin (batch No. 20200416-2, yellow bubble solid) prepared by the method described in patent document WO03099836A1 was added thereto, and the detection of impurities before purification was as shown in Table 6 below. Stirring until the system is dissolved. Another 50mL single-port bottle was taken, 9mL of purified water was added, stirring was started, 7.55g of sarcosine was added, and after stirring until the system was clear, the solution was added to the above dapagliflozin solution in absolute ethanol. The mixture is stirred and crystallized for 5 hours at the temperature of 20-30 ℃, filtered, and the collected solid is dried in vacuum for 6 hours at the temperature of 30-40 ℃ to obtain 25g of dapagliflozin sarcosine compound which is white solid with the batch number of 202000714-1, and the detection condition of impurities after purification is shown in the following table 6.

250ml (calculated as dapagliflozin sarcosine compound) of purified water was added to the reaction flask, stirring was turned on, dapagliflozin sarcosine compound was added, and 250ml (calculated as dapagliflozin sarcosine compound) of methyl tert-butyl ether was added. Stirring at 20-30 ℃ under the protection of nitrogen, standing, separating liquid, and collecting an organic phase. The organic phase is washed twice with 250ml of purified water, and the organic phase is decompressed and desolventized after liquid separation, thus obtaining a foaming solid product. The solid product is dissolved by 3 times volume of methyl tertiary butyl ether, added into 150ml of n-heptane solution, white solid is separated out, filtered under the protection of nitrogen, filter cake is collected, and the obtained product is dried in vacuum at 40 ℃ to obtain 19.5g of free dapagliflozin product (yield is 80%).

TABLE 6

Comparative example 2:

to a 3000mL three-necked flask, 520mL of methyl tert-butyl ether was added, stirring was started, 30g of dapagliflozin crude product (batch No. 20200416-2, yellow bubble solid) prepared according to the method provided in patent document WO03099836A1 was added, the conditions of impurity detection before purification were shown in Table 6, stirring was carried out until the system was cleared, 5.58g of (S) -1, 2-propanediol, 1.32g of purified water, 0.30g of dapagliflozin (S) -propanediol monohydrate seed crystal was continuously added according to the method provided in patent document WO2008002824A1, stirring was carried out at 20℃to 30℃for 8 hours, 1030mL of cyclohexane was added, the collected solid was filtered, vacuum-dried at 20℃to 25℃for 4 hours, and the impurity detection results after purification were shown in Table 6, 23g of dapagliflozin (S) -propanediol monohydrate as a white solid.

Comparative example 3:

to a 500mL single-necked flask, 170mL of isopropyl alcohol was added, followed by 35g (HPLC: 87.87%) of dapagliflozin crude product prepared by the method provided in WO03099836A1, and the mixture was stirred until the system was clear.

17mL of purified water is added into a 1000mL three-mouth bottle, stirring is started, 19.71-g L-proline is added, the system is heated to 80 ℃, 170mL of isopropanol is added, then the dapagliflozin isopropanol solution is added under rapid stirring, the temperature is slowly reduced to room temperature, the solution is stirred and crystallized for 5 hours at 20-30 ℃ under heat preservation, filtration is carried out, 20mL of isopropanol and 20mL of n-hexane are used for washing filter cakes, and the collected solid is dried in vacuum at 30-40 ℃ for 4 hours, thus obtaining 33.21g of dapagliflozin double L-proline eutectic white solid (HPLC: 97.41%).

To a 250mL three-necked flask, 50mL of purified water and 150mL of methyl t-butyl ether were added, 25g of the dapagliflozin double L-proline co-crystal obtained above was added, the organic phase was separated by stirring, the organic phase was washed 3 times with 50mL of purified water, and the solvent was distilled off under reduced pressure to obtain 14.8g of dapagliflozin as an off-white foamy solid (HPLC: 98.27%).

Example 10: purification of dapagliflozin crude product

150mL of absolute ethanol was added to a 250mL three-necked flask, stirring was started, 30g of a crude dapagliflozin product (batch No. 20200602, yellow bubble solid) prepared by the method provided in patent document WO03099836A1 was added, and the system was stirred until the system was cleared, as shown in Table 7 below, as the detection of impurities before purification. Another 50mL single-port bottle was taken, 10mL of purified water was added, stirring was started, 9.79g of sarcosine was added, and after stirring until the system was clear, the solution was added to the above dapagliflozin solution in absolute ethanol. The mixture is stirred and crystallized for 5 hours at the temperature of 20-30 ℃, filtered, and the collected solid is dried in vacuum for 6 hours at the temperature of 30-40 ℃ to obtain 23g of dapagliflozin sarcosine compound which is white solid with the batch number of 202000714-2, and the detection condition of impurities is shown in the following table 7.

TABLE 7

Comparative example 4:

600mL of methyl tert-butyl ether was added to a 3000mL three-necked flask, stirring was started, and 30g (batch: 202000602, yellow bubble solid) of dapagliflozin crude product prepared by the method described in patent document WO03099836A1 was added thereto, and the detection of impurities before purification was shown in Table 7. Stirring until the system is clear, continuing to add 5.58g of (S) -1, 2-propanediol, 1.54g of purified water and 0.30g of dapagliflozin (S) -propanediol monohydrate seed crystal according to the method of the example 6 for preparing dapagliflozin (S) -propanediol monohydrate in the patent document WO2008002824A1, stirring and crystallizing for 8 hours at the temperature of 20-30 ℃, adding 1030mL of cyclohexane, filtering, and vacuum drying the collected solid at the temperature of 20-25 ℃ for 4 hours to obtain 24g of off-white solid reaching the dapagliflozin (S) -propanediol monohydrate, wherein the batch number is 202000714-5, and the impurity detection result after purification is shown in Table 7.

Example 11: purification of dapagliflozin crude product

150mL of absolute ethanol was added to a 250mL three-necked flask, stirring was started, 30g of a crude dapagliflozin product (batch No. 20200703-3, yellow bubble solid) prepared by the method provided in patent document WO03099836A1 was added, and the system was stirred until the system was dissolved, as shown in Table 8 below, before the impurities were detected. Another 50mL single-mouth bottle was taken, 10mL of purified water was added, stirring was started, 9.8g of sarcosine was added, and after stirring until the system was clear, the solution was added to the above dapagliflozin solution in absolute ethanol. The mixture is stirred and crystallized for 5 hours at the temperature of 20-30 ℃, filtered, and the collected solid is dried in vacuum for 6 hours at the temperature of 30-40 ℃ to obtain 21g of dapagliflozin sarcosine compound which is white solid with the batch number of 20200714-3, and the detection result of impurities after purification is shown in the following table 8.

TABLE 8

Comparative example 5:

600mL of methyl tert-butyl ether was added to a 3000mL three-necked flask, stirring was started, 30g of dapagliflozin crude product (batch No. 20200703-3, yellow bubble solid) prepared according to the method provided in patent document WO03099836A1 was added, the mixture was stirred until the system was clear, 5.58g of (S) -1, 2-propanediol, 1.54g of purified water and 0.30g of dapagliflozin (S) -propanediol monohydrate seed crystal were continuously added according to the method provided in patent document WO2008002824A1, stirring was carried out at 20℃to 30℃for 8 hours, 1030mL of cyclohexane was added, the collected solid was filtered, and vacuum-dried at 20℃to 25℃for 4 hours, to obtain 20g of dapagliflozin (S) -propanediol monohydrate as white solid, batch No. 20200714-6, and the purified impurity was detected as shown in Table 8.

Comparative example 6:

50mL of absolute ethanol was added to a 100mL three-necked flask, stirring was started, 10g of dapagliflozin (0.025 mol, HPLC: > 99%) was added, and the mixture was stirred until the system was clear. Another 10mL single-necked flask was taken, 2mL of purified water was added, stirring was started, 2.20-g L-alanine (0.025 mol) was added, stirring was performed until the system was clear, and the solution was added to the above dapagliflozin ethanol solution. Stirring for 15 hours at-10℃to 10℃with no precipitation of solids, distilling off the solvent under reduced pressure to give a white solid, measuring the melting point to be about 70℃and having slightly non-melting particles, confirming that no eutectic is formed as a physical mixture of dapagliflozin and L-alanine.

Example 12: preparation of canagliflozin-sarcosine co-crystal

To a 100mL three-necked flask, 25mL of absolute ethanol was added, stirring was started, 5.00g of canagliflozin (HPLC: > 99%) was added, and the mixture was stirred until the system was clear. Another 10mL single-port bottle was taken, 3mL of purified water was added, stirring was started, 1.00g of sarcosine was added, and after stirring until the system was clear, the solution was added to the above-mentioned ethanol solution of canagliflozin. Crystallization is carried out for 5 hours at 20-30 ℃ with stirring, filtration is carried out, and the collected solid is dried in vacuum for 6 hours at 30-40 ℃ to obtain 5.16g white solid (HPLC: 99.96%).

By X-ray powder diffraction pattern (XRPD), single crystal diffraction, infrared spectrum analysis pattern (IR), differential scanning calorimetric pattern (DSC), thermogram (TGA), nuclear magnetic resonance hydrogen spectrogram ¹ H-NMR), see fig. 6-10, confirming that the white solid is a canagliflozin-sarcosine co-crystal.

¹ HNMR(600MHz,MeOD)δ7.535-7.511(m，2H)，7.306(s，1H)7.241-7.228(d，1H，J＝7.8Hz)，7.161-7.148(d，1H，J＝7.8Hz)，7.102-7.096(d，1H，J＝3.6Hz)，7.072-7.043(t，2H，J＝9.0Hz)，6.697-6.691(d，1H，J＝3.6Hz)，4.174-4.098(m，3H)，3.887-3.868(d，1H，J＝11.4Hz)，3.709-3.680(dd，1H，J＝12.0，5.4Hz)，3.487-3.465(m，3H)，3.429-3.368(m，3H)，2.663(s，3H)，2.294(s，3H)。

According to ¹ HNMR spectrum data shows that the molar ratio of canagliflozin to sarcosine is 1:1.

The canagliflozin-sarcosine co-crystals, using an X-ray powder diffraction (XRPD) pattern of Cu-K alpha radiation expressed in terms of diffraction angle 2θ, have characteristic peaks and their relative intensities as shown in table 9 below, with diffraction angle 2θ error of ± 0.2 °:

TABLE 9 Crgliflozin-sarcosine eutectic XRPD characteristic peaks and relative intensities thereof

No.	Diffraction angle 2 theta	D	I/Io	No.	Diffraction angle 2 theta	D	I/Io
1	3.625	24.3531	67.7	11	20.296	4.3719	80.1
2	7.096	12.4468	26.4	12	20.597	4.3086	24.2
3	10.595	8.3430	62.8	13	21.110	4.2051	47.2
4	14.077	6.2861	22.7	14	22.062	4.0257	20.2
5	15.933	5.5579	13.0	15	22.887	3.8824	77.0
6	16.768	5.2829	47.7	16	25.414	3.5018	29.1
7	17.338	5.1104	42.7	17	27.544	3.2356	18.5
8	18.322	4.8381	31.2	18	28.287	3.1523	45.7
9	18.791	4.7184	100	19	33.763	2.6525	5.1
10	19.603	4.5248	48.2	--	--	---	-

The infrared absorption (IR) spectrum of the canagliflozin-sarcosine eutectic has absorption peaks at the following positions measured by KBr tabletting: 3543.82 + -10 cm ^-1 、3153.82±10cm ^-1 、2690.68±5cm ^-1 、2603.06±5cm ^-1 、2419.53±5cm ^-1 、1596.33±5cm ^-1 、1507.48±5cm ^-1 、1411.84±5cm ^-1 、1382.27±5cm ^-1 、1321.84±5cm ^-1 、1233.44±5cm ^-1 、1086.11±5cm ^-1 、1062.19±5cm ^-1 、829.61±5cm ^-1 、799.52±5cm ^-1 、537.55±5cm ^-1 。

The Differential Scanning Calorimeter (DSC) spectrum of the canagliflozin-sarcosine eutectic has an endothermic peak in the range of 160.0-180.0 ℃, the peak value is 179.5 ℃, and the melting temperature is the melting temperature.

Thermal Gravimetric Analysis (TGA) profile of canagliflozin-sarcosine co-crystals with no significant weight loss before 150 ℃; the TGA-DTA diagram has a wider endothermic peak in the range of 190-230 ℃.

Example 13

Culturing to obtain single crystal reaching gliflozin-sarcosine eutectic in an ethanol/water mixed solvent system by using a solvent slow volatilization method, and carrying out X-ray single crystal diffraction characterization on the single crystal:

the single crystal structure of dapagliflozin-sarcosine co-crystal is shown in fig. 11:

TABLE 10 Single Crystal Structure data and Structure refinement parameters

TABLE 11 hierarchical atomic coordinates

Atoms	X	Y	Z
Cl1	-7182(3)	-8107.4(19)	-647.6(3)
O1	-8732(8)	-155(5)	-162.7(9)
O8	-3180(7)	101(6)	-1868.7(9)
O3	-1237(7)	-3339(5)	-1524.7(9)
O2	-4307(8)	-6295(5)	-1930.6(8)
O7	-4744(8)	-1431(5)	-1541.5(10)
O4	1151(8)	-3283(9)	-2038.1(10)
O6	-2966(10)	-8370(7)	-2359.5(10)
C16	-7078(12)	-2509(8)	-346.7(13)
C17	-8771(11)	-1394(7)	-337.2(12)
C22	-4852(11)	-408(7)	-1724.2(13)
C7	-4751(11)	-5779(7)	-1444.3(12)
N1	-9023(9)	-493(7)	-1629.0(11)
C8	-3640(11)	-7062(7)	-1341.1(12)
C21	-7086(13)	1522(7)	174.5(13)
C15	-7308(11)	-3743(7)	-534.5(13)
C20	-6743(12)	55(8)	14.4(13)
C6	-4063(11)	-5096(7)	-1725.4(12)
C5	-1605(10)	-4478(7)	-1734.1(12)
C11	-7407(11)	-5855(7)	-1044.3(11)
C2	-3907(11)	-5779(8)	-2214.4(12)
C18	-10675(11)	-1530(8)	-514.6(13)
O5	-1153(9)	-4729(11)	-2528.7(10)

Atoms	X	Y	Z
C14	-9196(11)	-3877(8)	-711.7(12)
C23	-7161(11)	287(7)	-1788.6(13)
C9	-4365(11)	-7751(7)	-1091.8(12)
C10	-6252(11)	-7149(8)	-949.1(12)
C13	-9564(11)	-5221(7)	-907.3(12)
C4	-1128(12)	-3904(10)	-2033.6(13)
C19	-10885(11)	-2734(8)	-699.1(12)
C12	-6588(10)	-5163(7)	-1294.8(12)
C3	-1454(11)	-5194(11)	-2240.6(13)
C24	-9140(11)	-136(9)	-1319.1(14)
C1	-4458(13)	-7105(9)	-2405.5(13)
H3	-2397	-2776	-1514
H4	1366	-2823	-2192
H6	-3222	-8744	-2199
H16	-5773	-2436	-227
H1C	-8841	-1515	-1649
H1D	-10389	-240	-1709
H8	-2372	-7473	-1442
H21A	-7245	2364	40
H21B	-5760	1707	297
H21C	-8475	1450	290
H15	-6147	-4499	-540
H20A	-5340	112	-103
H20B	-6584	-806	148
H6A	-5150	-4256	-1774
H5	-535	-5340	-1695
H2	-4989	-4927	-2257
H18	-11845	-780	-508
H5A	161	-4354	-2549
H23A	-7139	1378	-1737
H23B	-7469	209	-1994
H9	-3588	-8618	-1020
H13A	-10638	-4912	-1059

Atoms	X	Y	Z
H13B	-10309	-6046	-798
H4A	-2244	-3077	-2080
H19	-12186	-2796	-820
H12	-7302	-4262	-1362
H3A	-355	-6032	-2195
H24A	-9248	968	-1293
H24B	-7756	-516	-1225
H24C	-10493	-626	-1236
H1A	-6061	-7429	-2372
H1B	-4333	-6775	-2606

Analysis result

1) The X-ray single crystal diffraction characterization and the structural analysis show that the crystal belongs to an orthorhombic system, a space group P212121 and the unit cell parameters thereof are as follows:

table 12 cell parameters

2) The parameter of absolute configuration, flack parameter, was 0.03, less than 0.2, and the chiral center absolute configuration of dapagliflozin sarcosine co-crystal was determined by single crystal structural analysis as follows:

3) The molecular ellipsometry can determine that the components are dapagliflozin and sarcosine in a ratio of 1:1.

4) The following groups are bonded by hydrogen bonds and become eutectic.

Experimental example 1: dapagliflozin-sarcosine eutectic and dapagliflozin (S) -propylene glycol monohydrate influence factor comparison experiment

Dapagliflozin-sarcosine eutectic and dapagliflozin (S) -propylene glycol monohydrate are used as influence factor experiments, materials of the two crystal forms are respectively exposed for 10 days under the conditions of high temperature 60+/-2 ℃, illumination 4500 LX+/-500 LX and high humidity RH75% +/-5%, physical stability and chemical stability of the two crystal forms are examined, and the results are shown in the following tables 13, 14 and 15:

TABLE 13 results of co-crystal influencing factors of Glauber's net and sarcosine

TABLE 14 results of influence factors of dapagliflozin (S) -propylene glycol monohydrate

Table 15 dapagliflozin-sarcosine co-crystal influencing factor 10 day form stability

According to the experimental results of the influence factors, the number of detected substances and the total impurity detection amount of dapagliflozin and sarcosine eutectic are not increased under all experimental conditions, but the number of impurities and the total impurity detection amount of dapagliflozin (S) -propylene glycol monohydrate are increased at the high temperature of 60 ℃, the number of impurities is increased from 6 to 13 in 0 days, and the number of impurities and the detection amount of the dapagliflozin and sarcosine eutectic detected under all conditions of 10 days are consistent with 0 days, so that the dapagliflozin and sarcosine eutectic does not show a change trend. And (3) examining the XRPD and 0-day XRPD results of the 10-day sample under each condition of the influence factors, wherein the dapagliflozin eutectic is examined for 10 days under each condition of the influence factors, and the crystal form is unchanged.

Experimental example 2: comparative results of the effect factors of the co-crystal and sarcosine of canagliflozin and the hemihydrate of canagliflozin

The effect factor experiment is carried out on the canagliflozin-sarcosine eutectic and canagliflozin hemihydrate (original ground crystal form), materials of the two crystal forms are respectively exposed for 10 days under the conditions of high temperature 60+/-2 ℃, illumination 4500 LX+/-500 LX and high humidity RH75% +/-5%, and the physical stability and chemical stability of the two crystal forms are examined, and the results are shown in the following tables 16 and 17:

TABLE 16 comparative results of effect factors of canagliflozin-sarcosine co-crystal and canagliflozin hemihydrate

According to the experimental results of the influence factors, the canagliflozin-sarcosine eutectic is exposed for 10 days under the conditions of high temperature of 60 ℃ and high humidity of 75 percent, and compared with the detection results of 0 days, the number of the detected impurities and the detection amount do not show an increasing trend; placing under 5000LX illumination for 10 days, wherein 2 newly added impurities are detected by the canagliflozin eutectic, but the maximum detection amount is about 0.05%; the number of detected impurities is increased by 1 under the illumination condition of the hemihydrate; the chemical stability of the canagliflozin-sarcosine co-crystal is not poor Yu Kage columns of net hemihydrate. The weight of the canagliflozin hemihydrate is reduced by about 1.0% when the canagliflozin hemihydrate is exposed at the high temperature of 60 ℃ for 10 days, the weight of the canagliflozin hemihydrate is increased by about 2.1% when the canagliflozin sarcosine eutectic is exposed at the high temperature of 60 ℃ and the high humidity of 75%, and the weight increase are basically unchanged, so that the crystal form physical stability of the canagliflozin-sarcosine eutectic prepared by the invention is superior to that of the canagliflozin hemihydrate.

TABLE 17 Canagliflozin sarcosine co-crystal influencing factor 10 day Crystal form stability

And (3) examining the XRPD and 0-day XRPD results of the sample for 10 days under each condition of the influence factors, wherein the characteristic diffraction angle of the canagliflozin eutectic is basically consistent with that of the 0-day detection result after being examined for 10 days under each condition of the influence factors, so that the crystal form is unchanged.

Experimental example 3: dapagliflozin-sarcosine eutectic hygroscopicity experiment

The dapagliflozin-sarcosine co-crystal was subjected to a hygroscopicity test, and the results are shown in table 18 below:

TABLE 18 results of experiments on the wettability of dapagliflozin-sarcosine eutectic

From the above data, dapagliflozin-sarcosine co-crystals do not have hygroscopicity.

Experimental example 4: sieving comparison

A round sieve with the diameter of 20cm and the mesh number of 80 meshes is adopted, an electric oscillating sieve of 8411 type (Shaoxing city Yu area and Su Yu state geotechnical instruments factory) is adopted, the rotating speed is 1400 revolutions per minute, 30g of dapagliflozin-sarcosine eutectic crystal and dapagliflozin (S) propylene glycol compound hydrate are respectively sieved, the time is 20 minutes, the materials in a tray below the sieve are collected, the phenomenon after sieving is observed, the residue condition of the materials on the sieve and the condition of large particles are included, the adsorption condition of the materials by the sieve is observed, and the result is shown in the following table 19:

table 19 comparison of sieving results

The above yield results were subjected to one-way analysis of variance, and the results are shown in table 20 below:

TABLE 20 analysis of the sieving yield variance results

Source of discrepancy	SS	df	MS	F	P-value
Inter-group	51.627	1	51.627	20.85	0.01

From the above data, the two materials were screened for yield anova structure, which was significantly different.

According to the sieving yield and the observed phenomenon after sieving, the dapagliflozin-sarcosine eutectic powder state is more beneficial to sieving, has small sieving loss and high yield, and basically has no electrostatic effect.

Experimental example 5: dapagliflozin-sarcosine co-crystal and dapagliflozin- (S) propylene glycol monohydrate particle size and flowability comparison

The laser particle size analyzer is used for detecting the particle size distribution of dapagliflozin and sarcosine eutectic and dapagliflozin- (S) -propylene glycol monohydrate by taking water as a dispersing agent. And (3) respectively measuring the angle of repose, the bulk density and the tap density of dapagliflozin-sarcosine eutectic and dapagliflozin- (S) -propylene glycol monohydrate by adopting a BT-1000 powder comprehensive property tester. The results are shown in tables 21 and 22 below:

TABLE 21 particle size detection results

Name of product	D10 (micron)	D50 (micron)	D90 (micron)
Dapagliflozin-sarcosine co-crystal	5.151	24.83	89.93
Dapagliflozin- (S) -propylene glycol monohydrate	2.492	61.55	256.2

From the above results, dapagliflozin co-crystal D90 is 89.93 μm respectively, and the particle size distribution is favorable for uniform mixing in the preparation process of the pharmaceutical composition, especially for small-sized preparations.

TABLE 22 flowability test results

Name of product	Angle of repose (°)	Bulk density (g/ml)	Tap Density (g/ml)
Dapagliflozin-sarcosine co-crystal	40	0.40	0.58
Dapagliflozin- (S) -propylene glycol monohydrate	48	0.35	0.53

From the above results, it was found that the bulk density and tap density of dapagliflozin-sarcosine co-crystal and dapagliflozin- (S) -propylene glycol monohydrate were slightly different, the angle of repose of dapagliflozin-sarcosine co-crystal was smaller, and the fluidity was superior to dapagliflozin- (S) -propylene glycol monohydrate.

Experimental example 6: comparison of mixing difficulty in preparation production process

The prescription materials shown in table 20 were mixed by using an FH laboratory mixer (0.5L hopper) at a rotation speed of 8rpm for 30min, 5 points were taken at different positions in the hopper to detect the main drug content, RSD was calculated, and the mixing difficulty was evaluated, and the results are shown in Table 23.

Table 23 prescription composition

Prescription composition	Dosage (g)
Main medicine (charging quantity in dapagliflozin)	10.0
Microcrystalline cellulose	171.5
Anhydrous lactose	50.0
Crosslinked povidone	10.0
Croscarmellose sodium	4.0
Silica dioxide	3.75

Magnesium stearate

2.5

TABLE 24 flowability test results

The above results show that the same formulation and the same mixing process are adopted, the formulation with dapagliflozin and sarcosine eutectic as main drugs has the best mixing uniformity, and the preparation production requirement is met. Dapagliflozin- (S) -propylene glycol monohydrate is easy to agglomerate and is difficult to mix uniformly.

Experimental example 7: examination of the solubility of dapagliflozin-sarcosine co-crystals and Canagliflozin-sarcosine co-crystals in buffer salt solutions of different pH values

Taking dapagliflozin and sarcosine eutectic and proper amounts of dapagliflozin and sarcosine eutectic, respectively adding into pH 1.0 hydrochloric acid, pH 3.0 citrate buffer, pH 4.0 acetate buffer, pH 6.8 phosphate buffer, water, artificial gastric juice and artificial intestinal juice, shaking in a constant temperature air bath table at 25 ℃, respectively centrifuging for 10 minutes at 12 hours, 24 hours and 13000rpm/min, and taking supernatant for analysis, wherein the results are shown in the following table 25:

table 25 solubility examination (25 ℃ C.)

According to the measurement result, although the melting point of dapagliflozin-sarcosine eutectic is higher than that of dapagliflozin- (S) -propylene glycol monohydrate, the solubility in different pH media is still between 1.4 and 1.6mg/ml, and is consistent with the solubility of dapagliflozin- (S) -propylene glycol monohydrate reported in the literature; according to the solubility of the canagliflozin-sarcosine eutectic in different pH media, the solubility is about 30 mug/ml, and the canagliflozin-sarcosine eutectic is almost insoluble. According to the Japanese IF file of canagliflozin, the canagliflozin hemihydrate is almost insoluble in water (1 g is insoluble in 10000 ml), and the solubility of the two in aqueous solutions is substantially consistent. However, according to the physicochemical properties of the canagliflozin eutectic and the physical stability of the crystal forms, the canagliflozin eutectic is more suitable for the industrialized mass production of the preparation composition.

The above description of the embodiments is only for aiding in the understanding of the method of the present invention and its core ideas. It should be noted that it will be apparent to those skilled in the art that various modifications and adaptations of the invention can be made without departing from the principles of the invention and these modifications and adaptations are intended to be within the scope of the invention as defined in the following claims.

Claims

SGLT-2 inhibitor sarcosine co-crystal.
The SGLT-2 inhibitor-sarcosine co-crystal according to claim 1, wherein the X-ray powder diffraction pattern has diffraction peaks at 2Θ±0.2° positions of 10.6±0.2°, 19.6±0.2°, 22.1±0.2°, 33.6±0.2°.
SGLT-2 inhibitor-sarcosine co-crystal according to claim 1, characterized by having characteristic absorption peaks in the infrared spectrum at least at the following positions: 3540+ -10 cm ^-1 、2691±5cm ^-1 、2603±5cm ^-1 、2420±5cm ^-1 。
The SGLT-2 inhibitor sarcosine co-crystal of claim 1 wherein the thermogravimetric analysis has a broad endothermic peak at 190 ℃ to 230 ℃.
The SGLT-2 inhibitor-sarcosine co-crystal according to claim 1, wherein the nuclear magnetic resonance hydrogen spectrum (HNMR, 600MHz,MeOD) has-CH in the chemical shift range of 2.4 to 3.2ppm in addition to the formants having the SGLT-2 inhibitor structure ₃ Has a peak of-CH in the range of 3.0-4.0ppm ₂ -a peak.
The SGLT-2 inhibitor-sarcosine co-crystal of claim 1, wherein the SGLT-2 inhibitor is any one of dapagliflozin, engagliflozin, canagliflozin, tolagliflozin, isgliflozin, lu Gelie, sogliflozin, canagliflozin, begliflozin, exeagliflozin, elgliflozin, hengliflozin, regagliflozin, tagliflozin, valagliflozin, or a structure represented by formula a:
the SGLT-2 inhibitor-sarcosine co-crystal of claim 1 wherein the SGLT-2 inhibitor-sarcosine co-crystal is free of more than 0.1% of individual impurities.
The SGLT-2 inhibitor-sarcosine co-crystal according to claim 1, having a structure represented by formula i:

wherein R is ₁ 、R ₂ 、R ₃ 、R ₄ The ranges of X are shown in the following table:
the SGLT-2 inhibitor/sarcosine co-crystal according to claim 1, having a structure represented by formula ii:

wherein R is ₅ Is Cl, R ₆ Selected from any one of the following structures:

or R is ₅ Is methyl, R ₆ Is that

Or R is ₅ Is F, R ₆ Is that
The SGLT-2 inhibitor-sarcosine co-crystal of claim 1, wherein the SGLT-2 inhibitor to sarcosine molar ratio is 1: (0.5 to 5.0), in some embodiments, the SGLT-2 inhibitor to sarcosine molar ratio is preferably 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:0.95, 1:1.0, 1:1.05, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9 or 1:2.0; more preferred SGLT-2 inhibitors are present in a molar ratio to sarcosine of 1:0.8, 1:0.9, 1:0.95, 1:1.0, 1:1.05, 1:1.1, 1:1.2 or 1:1.3, 1:1.4 or 1:1.5.
The SGLT-2 inhibitor-sarcosine co-crystal of claim 1, wherein the SGLT-2 inhibitor is dapagliflozin, and wherein the SGLT-2 inhibitor-sarcosine co-crystal X-ray powder diffraction pattern has diffraction peaks at 2Θ±0.2° positions, the 2Θ±0.2° being 3.8±0.2°, 10.6±0.2°, 13.7±0.2 °, 17.0±0.2 °, 18.0±0.2 °, 18.6±0.2 °, 19.6±0.2 °, 20.1±0.2 °, 21.4±0.2 °, 22.1±0.2 °, 23.0.2 °, 25.4±0.2°, 27.6±0.2°, 33.6±0.2°.
The SGLT-2 inhibitor-sarcosine co-crystal according to claim 11, wherein single crystals reaching the gliflozin-sarcosine co-crystal are grown in an ethanol/water mixed solvent system by a slow solvent evaporation method, and the single crystals have the following unit cell parameters:

unit cell size:

α＝90°、β＝90°、γ＝90°

space group = P212121

Molecule/asymmetric unit=4

Wherein the measurement of the single crystal structure of the co-crystal is performed at t=112 and its hierarchical atomic coordinates are listed in table 11.
The SGLT-2 inhibitor-sarcosine co-crystal of claim 11 having an infrared spectrum with characteristic absorption peaks at the following positions: 3543.67 + -10 cm ^-1 、3161.94±10cm ^-1 、2690.95±5cm ^-1 、2603.78±5cm ^-1 、2419.39±5cm ^-1 、2360.665cm ^-1 、1599.61±5cm ^-1 、1510.92±5cm ^-1 、1291.19±5cm ^-1 、1045.97cm ^-1 。
The SGLT-2 inhibitor sarcosine co-crystal of claim 11 having an endothermic peak in the DSC profile at 140.0 ℃ to 155.0 ℃ at 149.0 ℃ and a melting temperature.
The SGLT-2 inhibitor sarcosine co-crystal of claim 11 having no significant weight loss before 150 ℃ in the TGA-DTA spectrum, having a broad endothermic peak in the range of 190 ℃ to 230 ℃.
The SGLT-2 inhibitor sarcosine co-crystal of claim 11 having a formant at a position below the nuclear magnetic resonance hydrogen spectrum: HNMR (600 mhz, meod) delta 7.343-7.329 (d, 1H, j=8.4 Hz), 7.315-7.312 (d, 1H, j=1.8 Hz) 7.276-7.259 (dd, 1H, j=8.4, 1.8 Hz), 7.091-7.077 (d, 2H, j=8.4 Hz), 6.795-6.781 (d, 2H, j=8.4 Hz), 4.089-4.073 (d, 1H, j=9.6 Hz), 4.055-3.977 (q, 2H, j=15.0 Hz), 3.994-3.959 (q, 2H, j=7.2 Hz), 3.876-3.854 (dd, 1H, j=12.0, 1.8 Hz), 3.467 (s, 2H) 3.461-3.431 (m, 1H), 37-3.409.370 (m, 37H), 37-37H (37.37H, 37-37 Hz).
The SGLT-2 inhibitor-sarcosine co-crystal of claim 1, wherein the SGLT-2 inhibitor is canagliflozin, and wherein the SGLT-2 inhibitor-sarcosine co-crystal X-ray powder diffraction pattern has diffraction peaks at 2Θ±0.2° positions of 3.6±0.2°, 7.1±0.2°, 10.6±0.2°, 14.1±0.2°, 16.8±0.2°, 17.3±0.2 °, 18.3±0.2 °, 18.8±0.2 °, 19.6±0.2 °, 20.3±0.2 °, 21.1±0.2°, 22.1±0.2°, 22.9±0.2°, 25.4±0.2°, 28.2±0.2°, 33.6±0.2°.
The SGLT-2 inhibitor-sarcosine co-crystal of claim 17 having an infrared spectrum with characteristic absorption peaks at the following positions: 3543.82 + -10 cm ^-1 、3153.82±10cm ^-1 、2690.68±5cm ^-1 、2603.06±5cm ^-1 、2419.53±5cm ^-1 、1596.33±5cm ^-1 、1507.48±5cm ^-1 、1086.11±5cm ^-1 、1062.19±5cm ^-1 、829.61±5cm ^-1 。
The SGLT-2 inhibitor/sarcosine co-crystal of claim 17 having an endothermic peak in the DSC profile at 160.0 ℃ to 180.0 ℃ at 179.5 ℃ and a melting temperature.
The SGLT-2 inhibitor-sarcosine co-crystal of claim 17 wherein said co-crystal has no significant weight loss in the TGA-DTA spectrum before 150 ℃ and has a broad endothermic peak in the range of 190 ℃ to 230 ℃.
The SGLT-2 inhibitor-sarcosine co-crystal according to claim 1, wherein the co-crystal has formants at the following positions in the nuclear magnetic resonance hydrogen spectrum HNMR (600 mhz, meod): delta 7.535-7.511 (m, 2H), 7.306 (s, 1H) 7.241-7.228 (d, 1H, j=7.8 Hz), 7.161-7.148 (d, 1H, j=7.8 Hz), 7.102-7.096 (d, 1H, j=3.6 Hz), 7.072-7.043 (t, 2H, j=9.0 Hz), 6.697-6.691 (d, 1H, j=3.6 Hz), 4.174-4.098 (m, 3H), 3.887-3.868 (d, 1H, j=11.4 Hz), 3.709-3.680 (dd, 1H, j=12.0, 5.4 Hz), 3.487-3.465 (m, 3H), 3.429-3.368 (m, 3H), 2.663 (s, 3H), 2.294 (s, 3H).
The preparation method of the SGLT-2 inhibitor and sarcosine co-crystal comprises the following steps:

mixing the SGLT-2 inhibitor solution and the sarcosine solution, standing for crystallization or cooling for crystallization, and carrying out solid-liquid separation to obtain the SGLT-2 inhibitor-sarcosine eutectic.
The method of claim 22, wherein the SGLT-2 inhibitor and the sarcosine are present in a molar ratio of 1: (0.5 to 5.0), in some embodiments, the SGLT-2 inhibitor to sarcosine molar ratio is preferably 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:0.95, 1:1.0, 1:1.05, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9 or 1:2.0.
the method of claim 22, wherein the solvent in the solution of the SGLT-2 inhibitor is selected from different single solvents or mixed solvents of C1-C10 alcohols, C3-C10 ketones, ethers, nitriles;

the solvent in the solution of sarcosine is selected from water.
The method according to claim 22, wherein the temperature of the standing crystallization or the cooling crystallization is-20 ℃ to 40 ℃; in some embodiments, the crystallization temperature is preferably: -15-35 ℃, 10-30 ℃, 5-30 ℃, 0-30 ℃, 5-30 ℃, 10-30 ℃, 15-30 ℃ and 20-30 ℃.
The method of claim 22, wherein the time for standing crystallization or cooling crystallization is 4-48 hours, preferably 4-24 hours, 4-16 hours, 4-12 hours, more preferably 8-12 hours.
A method for purifying a crude SGLT-2 inhibitor, comprising the steps of:

mixing the crude product of the SGLT-2 inhibitor with sarcosine, stirring at room temperature, and carrying out solid-liquid separation to obtain a sarcosine compound of the SGLT-2 inhibitor;

and dissociating sarcosinate of the SGLT-2 inhibitor to obtain a pure product of the SGLT-2 inhibitor in a free state.
The purification method according to claim 27, wherein the molar ratio of crude SGLT-2 inhibitor to sarcosine is 1: (0.5 to 5.0), in some embodiments, the molar ratio of crude SGLT-2 inhibitor to sarcosine is preferably 1:0.7, 1:0.8, 1:0.9, 1:0.95, 1:1.0, 1:1.05, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2.0 or 1:3.0; more preferred crude SGLT-2 inhibitor is present in a molar ratio to sarcosine of 1:1.0, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2.0 and 1:3.0.
The purification method of claim 27, further comprising:

directly preparing a final medicinal crystal form of the SGLT-2 inhibitor by taking the pure free SGLT-2 inhibitor as a raw material;

or the pure free SGLT-2 inhibitor is used as a raw material, and the final medicinal crystal form of the SGLT-2 inhibitor is prepared through recrystallization or eutectic mode.
The purification process according to claim 29, wherein the final pharmaceutically acceptable form of the SGLT-2 inhibitor is selected from the group consisting of pure SGLT-2 inhibitor, solvate, hydrate, solvate hydrate, co-crystal and double salt.
The purification method of claim 27, wherein the SGLT-2 inhibitor is pure in free form at a HPLC normalized purity of not less than 99%.
Pharmaceutical composition comprising the SGLT-2 inhibitor-sarcosine co-crystal according to any one of claims 1 to 21, or the SGLT-2 inhibitor-sarcosine co-crystal prepared by the method of preparation according to any one of claims 22 to 26, or the pure free form of the SGLT-2 inhibitor obtained by the purification method according to any one of claims 27 to 31, together with a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, vehicle or combination thereof.
Use of the SGLT-2 inhibitor-sarcosine co-crystal according to any one of claims 1 to 21, or the SGLT-2 inhibitor-sarcosine co-crystal prepared by the preparation method according to any one of claims 22 to 26, or the pure SGLT-2 inhibitor free form product obtained by the purification method according to any one of claims 27 to 31, or the pharmaceutical composition according to claim 32, for the preparation of a medicament for preventing, treating or alleviating cardiovascular and cerebrovascular diseases, diabetes and complications thereof, non-diabetes-induced nephropathy.
The use according to claim 33, wherein said diabetes and its complications are selected from one or more of essential hypertension, type 2 diabetes mellitus combined with hypertension, kidney disease combined with type 2 diabetes, kidney disease combined with hypertension and diabetes, kidney disease, type 1 diabetes, kidney disease of type 1 diabetes, liver fibrosis, insulin resistance, hyperglycemia, hyperinsulinemia, elevated blood levels of fatty acids or glycerol, hyperlipidemia, dyslipidemia, obesity.
Use of the SGLT-2 inhibitor-sarcosine co-crystal according to any one of claims 1 to 21, or the SGLT-2 inhibitor-sarcosine co-crystal prepared by the preparation method according to any one of claims 22 to 26, or the free pure SGLT-2 inhibitor product obtained by the purification method according to any one of claims 27 to 31, or the pharmaceutical composition according to claim 32, for the preparation of a medicament for lowering blood pressure.
A method for preventing, treating or alleviating diabetes mellitus and complications thereof, which comprises contacting a SGLT-2 inhibitor-sarcosine co-crystal according to any one of claims 1 to 21, or a SGLT-2 inhibitor-sarcosine co-crystal produced by the production method according to any one of claims 22 to 26, or a pure free form of the SGLT-2 inhibitor obtained by the purification method according to any one of claims 27 to 31, or a pharmaceutical composition according to claim 32, with a biological specimen.
A method for lowering blood pressure, comprising contacting the SGLT-2 inhibitor-sarcosine co-crystal according to any one of claims 1 to 21, or the SGLT-2 inhibitor-sarcosine co-crystal prepared by the preparation method according to any one of claims 22 to 26, or the pure SGLT-2 inhibitor free form product obtained by the purification method according to any one of claims 27 to 31, or the pharmaceutical composition according to claim 32, with a biological specimen.
A method for treating non-diabetes-induced nephropathy, comprising contacting the SGLT-2 inhibitor-sarcosine co-crystal according to any one of claims 1 to 21, or the SGLT-2 inhibitor-sarcosine co-crystal prepared by the preparation method according to any one of claims 22 to 26, or the pure free form of the SGLT-2 inhibitor obtained by the purification method according to any one of claims 27 to 31, or the pharmaceutical composition according to claim 32, with a biological specimen.